The miRCURY LNA SYBR Green PCR Kit contains a high-performance PCR master mix specifically designed for use with the miRCURY LNA miRNA PCR System. The miRCURY SYBR Green PCR Kit also features a built-in control for visual identification of correct template addition, as well as Q-Bond, an additive in the PCR buffer that enables short cycling steps without loss of PCR sensitivity and efficiency. The kit has been optimized for use with any real-time cycler. ROX Reference Dye is provided in a separate tube and can be added if your cycler requires ROX as a passive reference dye.

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The miRCURY LNA miRNA PCR System uses one single cDNA synthesis reaction for all amplifications, reducing pipetting and saving time and sample. Two LNA-enhanced miRNA-specific qPCR primers enable highly specific and sensitive amplification and single nucleotide discrimination. The fast and easy workflow takes only 3 hours with minimal hands-on steps.

The benefit of a universal reverse transcription step.

One universal RT reaction per RNA sample saves not only reagents and precious samples, but also your time.

Performance

Unmatched sensitivity

Universal reverse transcription combined with LNA-enhanced and Tm-normalized primers enables accurate and reliable quantification of individual miRNAs from as little as 1 pg total RNA (see figure Accurate quantitation from down to 1 pg total RNA starting material). With exceptional sensitivity and extremely low background, you can accurately quantify very low levels of miRNA without pre-amplification. This makes the miRCURY LNA miRNA PCR System suitable for all types of samples, especially those with low RNA content, such as serum, plasma and other biofluids.

Principle

High-performance reagents for quantitative, real-time RT-PCR

miRCURY LNA RT Kit
This kit includes all the necessary reagents to perform fast and convenient miRNA polyadenylation and reverse transcription in a single reaction step. Using a single, universal RT reaction as the template for subsequent PCR amplifications greatly simplifies the procedure compared to systems that require miRNA-specific, first-strand synthesis (see figure The benefit of a universal reverse transcription step). The UniSp6 RNA spike-in can be used with the control assay – UniSp6 miRCURY LNA miRNA PCR Assay (cat. no. YP00203954) – as a reverse transcription positive control and/or as an inter-plate calibrator in our miRNA PCR panels.

miRCURY LNA SYBR Green PCR Kit
This kit contains the high-performance miRCURY SYBR Green Master Mix for quantitative real-time PCR amplification. The detection is based on SYBR Green, which allows quality control of the resulting PCR amplicon by melting curve analysis. LNA-enhanced primer sets require specific PCR reaction conditions for optimal performance. The kit also includes a control primer set designed for amplification of the synthetic RNA spike-in provided in the miRCURY LNA RT Kit. Refer to the miRCURY LNA RT Kit Handbook for details. The miRCURY LNA miRNA PCR Assays and Panels have been validated using the miRCURY SYBR Green Master Mix; use of other reagents for qPCR will affect the results.

Procedure

The miRCURY LNA miRNA PCR Assay system is an miRNA-specific, LNA-based system designed for sensitive and accurate detection of miRNA by quantitative real-time PCR using SYBR Green. The first step of the procedure is universal reverse
transcription, followed by real-time PCR amplification with LNA-enhanced primers (see figure miRCURY LNA miRNA PCR System at a glance).

Applications

The miRCURY LNA SYBR Green PCR Kits are used as part of the miRCURY PCR System for: